• Title/Summary/Keyword: 차세대염기서열 분석

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Microbial Forensics: Comparison of MLVA Results According to NGS Methods, and Forensic DNA Analysis Using MLVA (미생물법의학: 차세대염기서열분석 방법에 따른 MLVA 결과 비교 및 이를 활용한 DNA 감식)

  • Hyeongseok Yun;Seungho Lee;Seunghyun Lim;Daesang Lee;Sehun Gu;Jungeun Kim;Juhwan Jeong;Seongjoo Kim;Gyeunghaeng Hur;Donghyun Song
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.4
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    • pp.507-515
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    • 2024
  • Microbial forensics is a scientific discipline for analyzing evidence related to biological crimes by identifying the origin of microorganisms. Multiple locus variable number tandem repeat analysis(MLVA) is one of the microbiological analysis methods used to specify subtypes within a species based on the number of tandem repeat in the genome, and advances in next generation sequencing(NGS) technology have enabled in silico anlysis of full-length whole genome sequences. In this paper, we analyzed unknown samples provided by Robert Koch Institute(RKI) through The United Nations Secretary-General's Mechanism(UNSGM)'s external quality assessment exercise(EQAE) project, which we officially participated in 2023. We confirmed that the 3 unknown samples were B. anthracis through nucleic acid isolation and genetic sequence analysis studies. MLVA results on 32 loci of B. anthracis were analysed by using genome sequences obtained from NGS(NextSeq and MinION) and Sanger sequencing. The MLVA typing using short-reads based NGS platform(NextSeq) showed a high probability of causing assembly error when a size of the tandem repeats was grater than 200 bp, while long-reads based NGS platform(MinION) showed higher accuracy than NextSeq, although insertion and deletion was observed. We also showed hybrid assembly can correct most indel error caused by MinION. Based on the MLVA results, genetic identification was performed compared to the 2,975 published MLVA databases of B. anthracis, and MLVA results of 10 strains were identical with 3 unkonwn samples. As a result of whole genome alignment of the 10 strains and 3 unknown samples, all samples were identified as B. anthracis strain A4564 which is associated with injectional anthrax isolates in heroin users.

Application of NGS Analysis for the Food Source of Bivalve (이매패류(Sinonovacula constricta) 먹이원 NGS 분석 적용에 대한 연구)

  • Heo, Yu-Ji;Jo, Hyunbin;Jung, Eunsong;Kim, Hyun-Woo
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.257-264
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    • 2021
  • In this study, we analyzed the food components in the release product that sampled Sinonovacula constricta from the foreshore littoral at Byeongnyang-myeon, Suncheon Bay. We used microscopy and next-generation sequencing (NGS) to evaluate the applicability of morphological and molecular methods to analyze release products. The higher species diversity observed in the NGS method is due to the different levels of species identification, as microscopy displays morphological and anatomical levels of plankton species identification in S. constrita. Moreover, NGS can identify the level of species in the organic matter by using the 18s_V9 primer.

Development of Chloroplast Genome-based Insertion/Deletion Markers in the Genus Broussonetia (닥나무 속 식물의 엽록체 유전체 기반 InDel 마커의 개발)

  • Eun Jee Lee;Yoon A Kim;Mi Sun Lee;Ju Hyeok Kim;Young Kyu Choi;Jung Sung Kim;Chang Seob Sin;Yi Lee
    • Korean Journal of Plant Resources
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    • v.36 no.4
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    • pp.290-298
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    • 2023
  • Several members of the genus Broussonetia are woody plants with high-quality cellulose fibers and are used to make a traditional type of Korean paper known as Hanji. Three of these species, Broussonetia kazinoki, Broussonetia monoica, and Broussonetia papyrifera, are found in the Korean Peninsula. Because it is challenging to distinguish different Broussonetia species based on morphology alone, we have developed a set of insertion/deletion (InDel) markers for genetic identification of these species. From twenty-two Broussonetia samples collected throughout Korea, we selected six for next-generation sequencing analysis. InDel marker candidates were identified by comparing this sequence information with the B. kazinoki chloroplast genome sequence. The marker candidates were used to screen the genomes of the twenty-two Broussonetia plants, and five useful chloroplast-based InDel markers were identified. Detailed genotyping using these five markers showed that the twenty-two plants of the genus Broussonetia could be clustered into five groups, verifying that the markers developed here can be used for breeding, identification, and analysis of species in the genus Broussonetia.

Development of SNP Molecular Marker for Red-fleshed Color Identification of Peach Genetic Resources (복숭아 유전자원의 적색 과육 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Nam, Eun Young;Cho, Kang Hee;Jun, Ji Hae;Chung, Kyeong Ho
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.303-311
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    • 2019
  • Various colors of fruit skin and flesh are the most popular commercial criteria for peach classification. In order to breed new red-fleshed peach cultivar, many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection markers to screen seedlings with target traits to increase breeding efficiency. For the comparison of transcription profiles in peach cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red-fleshed peach cultivar, 'Josanghyeoldo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two cultivars were selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNP) were screened from peach EST contigs by high resolution melting (HRM) analysis displayed specific difference between 8 red-fleshed peach cultivars and 24 white-fleshed peach cultivars. All 72 pairs of SNPs were discriminated and the HRM profiles of amplicons were established. In the study reported here, the development of SNP markers for distinguishing between red and white fleshed peach cultivars by HRM analysis offers the opportunity to use DNA markers. This SNP marker could be useful for peach marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach cultivars.

SNP Markers Useful for the Selection of Yellow-fleshed Peach Cultivar (황육계 복숭아 품종 선발용 SNP 마커)

  • Kim, Se Hee;Kwon, Jung-hyun;Cho, Kang Hee;Shin, Il Sheob;Jun, Ji Hae;Cho, Sang-Yun
    • Korean Journal of Plant Resources
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    • v.34 no.5
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    • pp.443-450
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    • 2021
  • Peach flesh color is commercially important criteria for classification and has implications for nutritional quality. To breed new yellow-fleshed peach cultivar many cross seedlings and generations should be maintained. Therefore it is necessary to develop early selection molecular markers for screening cross seedlings and germplasm with economically important traits to increase breeding efficiency. For the comparison of transcription profiles in peach varieties with a different flesh color expression, two cDNA libraries were constructed. Differences in gene expression between yellow-fleshed peach cultivar, 'Changhowon Hwangdo' and white-fleshed peach cultivar, 'Mibaekdo' were analyzed by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the two varieties was selected for nucleotide sequence determination and homology searches. Putative single nucleotide polymorphisms (SNPs) were screened from peach EST contigs by high resolution melting (HRM) analysis, SNP ID ppa002847m:cds and ppa002540m:cds displayed specific difference between 17 yellow-fleshed and 21 white-fleshed peach varieties. The SNP markers for distinguishing yellow and white fleshed peach varieties by HRM analysis offers the opportunity to use early selection. This SNP markers could be useful for marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in peach varieties.

Microbial community structure analysis from Jeju marine sediment (제주도 인근 해양퇴적물 내의 미생물 군집 구조분석)

  • Koh, Hyeon Woo;Rani, Sundas;Hwang, Han-Bit;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.375-379
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    • 2016
  • In this study, the structure and diversity of bacterial community were investigated in the surface and subsurface marine sediments using a NGS method (i.e. illumina sequencing technology). The bacterial community in the surface was distinct from that in the subsurface of marine sediment; with the exception of the phylum Proteobacteria, the relative abundance of Bacteroides phylum were higher in the surface than subsurface, whereas the sequences affiliated to the phyla Chloroflexi and Acidobacteria were relatively more copious in the subsurface than surface sediment. Moreover, interestingly, we observed that the phyla Nitrospinae and Nitrospirae contribute to nitrogen cycle in the marine sediment. This study may present the possibility for the presence of novel microorganisms as unexplored sources and provide basic information on the microbial community structure.

The Application of Genome Research to Development of Aquaculture (양식산업에 발전을 위한 유전체 분석 기술 적용)

  • Lee, Seung Jae;Kim, Jinmu;Choi, Eunkyung;Jo, Euna;Cho, Minjoo;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.2
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    • pp.47-57
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    • 2021
  • In the fishery industry, global aquaculture production has stagnated due to overfishing of aquatic products, restrictions between countries, and climate change. The aquaculture suggests the possibility of a blue revolution that can be expanded in a new way. The aquaculture industry now accounts for more than half of the fishery products from the sea as a raw material for seafood for human consumption. Various latest biological research methods are being applied for the development of a sustainable aquaculture industry. Genomics has made significant progress in recent years. Since the genome sequence of Atlantic cod was sequenced in 2011, the genomes of more species have been sequenced. The genome information is providing a more robust and productive knowledge base for the aquaculture industry, including breeding and breeding of superior traits, improving disease resistance quality, and optimizing aquaculture feed and feed methods. This review looked at the status of genome analysis technology and the current status of genome research of aquaculture species. The development of genome research technology and massive genomic information is important in solving the challenges of the aquaculture industry and will help sustainable fisheries and aquaculture.

Identification of Erysiphe izuensis on Rhododendron yedoense f. poukhanense in Korea Based on Morphological and Molecular Characteristics (형태 특징 및 분자 분석에 의한 산철쭉 흰가루병균 Erysiphe izuensis 동정)

  • Cho, Sung-Eun;Lee, Sang-Hyun;Lee, Sun-Keun;Seo, Sang-Tae;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.46 no.1
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    • pp.69-74
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    • 2018
  • The Rhododendron powdery mildew fungus Erysiphe izuensis has been recorded throughout Asia, including China, Japan, Korea, and Russia. In Korea, E. izuensis has been identified based on morphological characteristics of the anamorph found on introduced Rhododendron spp. We here describe the first identification of E. izuensis collected from Rhododendron yedoense f. poukhanense in Korea. Morphological characteristics of the teleomorph as well as internal transcribed spacer sequences of the species are provided. Moreover, we describe the characteristics of the primary conidia and conidial surface pattern of E. izuensis for the first time.

Investigation of microplastic biofilm communities originated from freshwater (미세플스틱 표면에 형성된 담수 유래 생물막 군집 고찰)

  • Choi, Woodan;Nguyen, Hien Thi;Kim, Eun-Ju;Cho, Kyungjin
    • Journal of Korean Society of Water and Wastewater
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    • v.36 no.2
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    • pp.97-106
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    • 2022
  • Recently microplastic (MP) biofilm is being attracted as an important environmental issue because it can act as a pollutant carrier in aqueous system. Therefore, this study investigated the MP biofilm communities originated from freshwater. The results showed the bacterial community structure of MP biofilm was distinctively different from the freshwater regardless of biofilm-forming condition and MP type. For MP biofilm communities exposed to raw freshwater, Solimonas variicoloris-like microbe, Frigidibacter albus-like microbe, Nitrospirillum amazonense-like microbe, and Pseudochroococcus couteii-like microbe became abundant, while Acinetobacter johnsonii, Macellibacteroides fermentans, and Sedimentibacter acidaminivorans-like microbe were found as major bacteria for MP biofilm communities exposed to organic rich condition. The results of this study suggest that the unique freshwater biofilm community could be formed on the MP surface.

Genomic epidemiology and surveillance of zoonotic viruses using targeted next-generation sequencing (표적화 차세대염기서열분석법을 이용한 인수공통 바이러스의 유전체 역학과 예찰)

  • Seonghyeon Lee;Seung-Hwan Baek;Shivani Rajoriya;Sara Puspareni;Won-Keun Kim
    • Korean Journal of Veterinary Service
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    • v.46 no.1
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    • pp.93-106
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    • 2023
  • Emerging and re-emerging zoonotic viruses become critical public health, economic, societal, and cultural burdens. The Coronavirus disease-19 (COVID-19) pandemic reveals needs for effective preparedness and responsiveness against the emergence of variants and the next virus outbreak. The targeted next-generation sequencing (NGS) significantly contributes to the acquisition of viral genome sequences directly from clinical specimens. Using this advanced NGS technology, the genomic epidemiology and surveillance play a critical role in identifying of infectious source and origin, tracking of transmission chains and virus evolution, and characterizing the virulence and developing of vaccines during the outbreak. In this review, we highlight the platforms and preparation of targeted NGS for the viral genomics. We also demonstrate the application of this strategy to take advantage of the responsiveness and prevention of emerging zoonotic viruses. This article provides broad and deep insights into the preparedness and responsiveness for the next zoonotic virus outbreak.